Bani Hani, Saja MS

Evans, Gethin H

Flint, Pamela

Graf, Brigitte A

E-space: Manchester Metropolitan University's Research Repository

Bani Hani, Saja MS and Flint, Pamela and Evans, Gethin H and Graf, BrigitteA (2020) Commonly used method for chylomicron isolation catches less than0.1% of chylomicrons from whole plasma. In: 13th European Nutrition Con-ference, FENS 2019, 15 October 2019 - 18 October 2019, Dublin, Republicof Ireland.Downloaded from: https://e-space.mmu.ac.uk/626382/Version: Published VersionPublisher: Cambridge University Press (CUP)DOI: https://doi.org/10.1017/S0029665120002311Please cite the published versionhttps://e-space.mmu.ac.ukThe 13th European Nutrition Conference, FENS 2019, was held at the Dublin Convention Centre, 15–18 October 2019Commonly used method for chylomicron isolation catches less than 0.1%of chylomicrons from whole plasmaSaja Bani Hani, Pamela Flint, Gethin Evans and Brigitte GrafManchester Metropolitan University, Manchester, United KingdomAbstractHealth effects of orally ingested bioactive compounds can only occur if bioactive molecules are absorbed and transported to targettissues. Paradoxically, many foods are rich in micronutrients but intestinal absorption is often limited. Immediately after absorption,lipid soluble nutrients are packaged into chylomicrons (CM), therefore quantification of the micronutrient content in CM has beenused as tool to evaluate bioavailability(1). For bioavailability studies the isolated CM fraction must not contain liver derived lipopro-teins (VLDL, LDL) since only intestine derived CM carry recently absorbed lipids and lipophilic micronutrients. As isolated CM canbe contaminated with liver derived lipoproteins, this study evaluated the purity of collected CM and VLDL fractions. Each CM con-tains one apolipoprotein B-48 (apoB-48) and each VLDL contains one apolipoprotein B-100 (apoB-100) on its surface(2) thereforepurity of CM and VLDL fractions was evaluated via the presence/absence of apoB-100 and apoB-48.CM and VLDL fractions were isolated as previously described(1,3)from whole plasma collected at 0, 2, 4 or 6 h after participantsconsumed a lipid (15.4g) and carotenoid (36mg) rich smoothie (480mL). Plasma density was adjusted to 1.1g/mL with KBr (0.14g/mL),placed in ultracentrifugation tubes (38 mL, thickwall) and overlaid with 3 solutions containing NaCl, KBr and Na-EDTA with densitiesof 1.020, 1.065 and 1.006g/mL.CM fraction was collected after ultracentrifugation at 25,000 rpm for 34min in a Beckman Coulter OptimaXE-90 with SW32Ti swinging bucket rotor. VLDL fraction was collected after additional 102min at 25,000 rpm.ELISA analysis revealed that apoB-48 and apoB-100 were present in both CM and VLDL fractions. Less than 0.1% of plasmaapoB-48 (< 98pmol/L) was present in CM or VLDL fraction. Less than 0.3% (< 2,425pmol/L) and 4% (< 54,124pmol/L) of plasmaapoB-100 was present in CM and VLDL fractions, respectively. Low recoveries of apoB-48 and apoB-100 in CM and VLDL fractionsuggests that ultracentrifugation neither concentrated nor isolated CM or VLDL. While ourapoB-48 and apoB-100 results agree withpreviously reported values in plasma, CM and VLDL(3,4), no previous study evaluated apoB-48 and apoB-100 recovery from plasma.Conflict of InterestThere is no conflict of interestReferences1. White W, Zhou Y, Crane, et al. (2017) Am J Clin Nutr 106, 1041–1051.2. Burnett J & Watts G (2008) Clin Chem 54, 782–784.3. Zhang Y, Zhou Y, Gribnau et al. (2014) Graduate Theses and Dissertations 13731.4. Masuda D, Sakai N, Sugimoto et al. (2011) J Atheroscler Thromb 18, 1062–1070.Proceedings of the Nutrition Society (2020), 79 (OCE2), E283 doi:10.1017/S0029665120002311ProceedingsoftheNutritionSocietyhttps://www.cambridge.org/core/terms. https://doi.org/10.1017/S0029665120002311Downloaded from https://www.cambridge.org/core. Manchester Metropolitan University, on 24 Aug 2020 at 12:31:08, subject to the Cambridge Core terms of use, available at

Commonly used method for chylomicron isolation catches less than 0.1% of chylomicrons from whole plasma.

https://e-space.mmu.ac.uk/626382/1/Bani%20Hani%20SMS%20Graf%20BAG%202019%20FENS%20abstract.pdf

Commonly used method for chylomicron isolation catches less than 0.1% of chylomicrons from whole plasma.

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